Takemoto Konosuke, Mikota Yuichi, Moriuchi Ryota, Yoneda Yuko, Kawai Shingo
Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka, 422-8529, Japan.
The United Graduate School of Agricultural Science, Gifu University, 1-1, Yanagido, Gifu-shi, 501-1193, Japan.
Heliyon. 2024 Mar 9;10(6):e27698. doi: 10.1016/j.heliyon.2024.e27698. eCollection 2024 Mar 30.
is an actinorhizal plant that coexists with the nitrogen-fixing actinomycete via nodules. It produces a variety of polyketides, including flavonoids, stilbenoids, and diarylheptanoids. These compounds have beneficial biological activities. Plant polyketides are produced by type III polyketide synthases (PKSIII). In this study, three PKSIIIs (AsPKSIII1, AsPKSIII2, and AsPKSIII3) predicted from next-generation sequencing analysis of seedling RNA were amplified and cloned. Phylogenetic tree analysis classified AsPKSIII2 and AsPKSIII3 into the chalcone synthase (CHS) group, whereas AsPKSIII1 was not classified into this group. We attempted to produce polyketides by adding cinnamic acid analogs to the culture medium of in which the respective PKSIII gene and the acetyl-CoA carboxylase (ACC) and 4-coumarate: CoA ligase (4CL) genes were simultaneously recombined. AsPKSIII1 is an enzyme that condensed only one molecule of malonyl-CoA to cinnamoyl-CoAs. In contrast, AsPKSIII2 and AsPKSIII3 produced chalcones as shown in a phylogenetic tree analysis, but also produced triketide pyrone. The ratio of these products differed between the two enzymes. We determined the gene and amino acid sequences as well as the substrate specificities of the two enzymes involved in flavonoid production and one enzyme potentially involved in diarylheptanoid production in .
是一种通过根瘤与固氮放线菌共生的放线菌根植物。它能产生多种聚酮化合物，包括黄酮类、芪类和二芳基庚烷类。这些化合物具有有益的生物活性。植物聚酮化合物由III型聚酮合酶（PKSIII）产生。在本研究中，从幼苗RNA的下一代测序分析预测的三种PKSIIIs（AsPKSIII1、AsPKSIII2和AsPKSIII3）被扩增并克隆。系统发育树分析将AsPKSIII2和AsPKSIII3归类为查尔酮合酶（CHS）组，而AsPKSIII1未被归类到该组。我们试图通过向分别同时重组了各自PKSIII基因、乙酰辅酶A羧化酶（ACC）和4-香豆酸：辅酶A连接酶（4CL）基因的培养基中添加肉桂酸类似物来生产聚酮化合物。AsPKSIII1是一种仅将一分子丙二酰辅酶A缩合到肉桂酰辅酶A上的酶。相比之下，AsPKSIII2和AsPKSIII3如系统发育树分析所示产生了查尔酮，但也产生了三酮吡喃。这两种酶产生的这些产物的比例不同。我们确定了参与黄酮类化合物生产的两种酶以及可能参与二芳基庚烷类化合物生产的一种酶的基因和氨基酸序列以及底物特异性。
